Alexa Fluor 488 anti-Vimentin Antibody anti-Vimentin

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Clone: O91D3 (See other available formats)
Regulatory Status: RUO
Other Names: CTRCT30, HEL 113, Epididymis Luminal Protein 113
Isotype: Mouse IgG2a, κ
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Product Citations: publications

O91D3_A488_Vimentin_Antibody_102720 — HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 0.5 µg/ml Alexa Fluor® 488 anti-Vimentin (clone O91D3) or Alexa Fluor® 488 Mouse IgG2a, κ Isotype Ctrl Antibody overnight at 4 degree. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.

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677809

25 µg

109€

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Description

Vimentin are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is a widely expressed and highly conserved 54 kD protein that is constitutively expressed in mesenchymal cells, endothelial cells lining blood vessels, renal tubular cells, macrophages, neutrophils, fibroblasts, and leukocytes^1,2. Vimentin is used as a marker of mesenchymal cells to distinguish them from epithelial cells³. Increased vimentin expression is frequently used as an EMT marker in cancer⁴. Autoantibodies to vimentin are commonly found in patients with autoimmune diseases such as Lupus⁵ and rheumatoid arthritis⁶, and also found after transplantation⁷.

Product Details

Technical Data Sheet (pdf)

Product Details

Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Full length human vimentin produced in E. coli.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: ICC - Quality tested
ICFC, IHC-P - Verified
SB - Community Verified
Recommended Usage: Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, the suggested use of this reagent is 0.2 - 2 µg per mL. For intracellular flow cytometry using our True-Phos™ Perm Buffer, the suggested use of this reagent is ≤ 1.0 µg per million cells in 100 µL volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.

Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser: Blue Laser (488 nm)
Application Notes: While this clone recognizes mouse Vimentin, we do not recommend its usage for western blot due to poor affinity of the antibody for the protein. Additional reported applications for the relevant formats include: spatial biology (IBEX)^1,2.
Additional Product Notes: For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.
RRID: AB_2650955 (BioLegend Cat. No. 677809)

Antigen Details

Structure: 466 amino acids with a predicted molecular weight of approximately 54 kD.
Distribution: Cytoplasm.
Function: Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
Interaction: HCV core protein, LGSN, SYNM, PLEC, SLC6A4, STK33, LARP6, RAB8B, TOR1A, TOR1AIP1, and BCAS3.
Cell Type: B cells, Mesenchymal Stem Cells, Neural Stem Cells, Neutrophils
Biology Area: Cell Adhesion, Cell Biology, Cell Motility/Cytoskeleton/Structure, Immunology, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family: Intermediate Filaments
Antigen References: 1. Kidd ME, et al. 2014. Am. J. Respir. Cell Mol. Biol. 50:1.
2. Fuchs E, et al. 1994. Annu. Rev. Biochem. 63:345.
3. Zeisberg M, et al. 2009. J. Clin. Invest. 119:1429.
4. Scanlon CS, et al. 2013. J. Dent. Res. 92:114.
5. Thebault S, et al. 2002. J. Immunol. 169:4046.
6. Vossenaar ER, et al. 2004. Arthritis Res. Ther. 6:R142.
7. Rose ML. 2013. Hum. Immunol. 74:1459.
Gene ID: 7431 View all products for this Gene ID
UniProt: View information about Vimentin on UniProt.org

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Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Are other fluorophores compatible with IBEX?: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
The same antibody works in one tissue type but not another. What is happening?: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
How can I be sure the staining I’m seeing in my tissue is real?: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All Vimentin Reagents Request Custom Conjugation

Description	Clone	Applications
Alexa Fluor® 594 anti-Vimentin	O91D3	ICC,IHC-P,SB
Purified anti-Vimentin	O91D3	WB,ICC,ICFC,IHC-P,IHC-F,SB
Direct-Blot™ HRP anti-Vimentin	O91D3	WB
Alexa Fluor® 647 anti-Vimentin	O91D3	ICC,IHC-P,SB
Alexa Fluor® 488 anti-Vimentin	O91D3	ICC,ICFC,IHC-P,SB
TotalSeq™-Bn1302 anti-Vimentin	O91D3	SB
PerCP/Cyanine5.5 anti-Vimentin	O91D3	ICFC,SB

Certificate of Analysis

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Go To Top Version: 6 Revision Date: 10.10.2024

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